17

Maintenance of Cannabis germplasm in the Vavilov Research Institute Gene Bank - 1996

Sofia Kutuzova ¹, Lyudmila Rumyantseva ¹, Sergey Grigoryev ¹ and Robert C. Clarke ²

¹ N. I. Vavilov All-Russian Research Institute of Plant Industry, St. Petersburg, Russia; ² International Hemp Association, Postbus 75007, 1070 AA Amsterdam, The Netherlands

        Kutuzova, Sofia, L. Rumyantseva, S. Grigoryev and R. C. Clarke 1997.  Maintenance of Cannabis germplasm in the Vavilov Research Institute Gene Bank - 1996.  Journal of the International Hemp Association 4(1): 17-21.  Ninety-six accessions of hemp were grown on isolated plots at the Pavlovsk, Yekaterinino, Kuban and Ustimovka Experiment Stations of the N. I. Vavilov All-Russian Research Institute of Plant Industry (VIR) and at the Penza Research Institute of Agriculture and the Donskoy Branch of the All-Russian Research Institute of Oil-bearing Crops.  Reproductions were received from only 74 accessions because six did not germinate, two perished during chemical weed control of the covering crop, and 14 were stolen.  In addition, small amounts of seed were received from 16 samples of monoecious hemp grown in glasshouses at the Pushkin Laboratories of the VIR.

Introduction
        The Cannabis collection preserved at the VIR consists of 496 accessions, representing the wide global diversity of this crop.  Current priorities for the conservation of Cannabis genetic diversity are the maintenance of the collection in viable condition through reproductions in the field, and placement of the reproduced accessions in long-term storage.  However, due to the worsening situation with respect to scientific research in Russia, implementation of these tasks becomes ever more difficult and expensive from year to year.
        The loss of southern experiment stations, where late-ripening thermophilous (heat-loving) hemp accessions had been maintained, led to attempts to reproduce these genotypes in more northern environments, where they sometimes failed to yield mature seed.  This produced the necessity to incorporate in this project, new institutions in southern regions with favorable climates, which have the possibility to grow out the collection materials on isolated field plots or under reliable artificial isolation.
        Incidents involving the stealing of plants became more frequent, indicating that the precautions taken were not successful.  An attempt to grow out several accessions at the Donskoy Branch of the All-Russian Institute of Oil-bearing Crops (Rostov Province) was futile due to these reasons.  Many more plant samples were lost at the Kuban and Yekaterinino Stations of the VIR.
        An experiment was performed at the Pushkin laboratories of the VIR to reproduce monoecious hemp accessions in glasshouses, but the possibilities were limited by the small number of boxes and small number of seeds planted.

Methods
        In 1996, participants in the VIR/IHA Cannabis Germplasm Preservation Project continued with their efforts to reproduce the next set of Cannabis collection accessions.  Planting was performed on isolated plots at the same stations as in the previous year: the Pavlovsk, Yekaterinino, Kuban and Ustimovka Experiment Stations of the VIR.  There was also an attempt to expand the scope of work by the incorporating new sites: the Penza Research Institute of Agriculture (Penza Province) and the All-Russian Institute of Oil-bearing Crops (Rostov Province), and by growing monoecious hemp accessions in the glasshouses of the Pushkin Laboratories of the VIR (near St. Petersburg).
        The methods applied to the isolated plots were the same as in previous years (Lemeshev et al. 1994, 1995 and Kutuzova et al. 1996): manual planting in rows with an interval of 20 cm between the rows, hoeing, weed control, top-dressing of young plants, eliminating atypical plants in the budding and flowering phases, and manual harvesting.
        In the glasshouse, monoecious hemp samples were grown in pots of five liter capacity.  Ten pots were used for each accession and three to five seeds were sown in each pot, with subsequent regular watering and weeding.   At the beginning of budding, the pots of each accession were placed in isolation boxes.  Varietal integrity was maintained by eliminating atypical off-type plants and dioecious individuals from the crop.  The female plants were periodically pollinated by shaking the males.

Results
        Weather conditions in 1996 were extremely unfavorable for hemp cultivation at all the locations selected for reproductions.  Spring droughts and arid winds in the southern and central regions of the country (including Kuban, Ustimovka, Yekaterinino and Donskoy Branch) caused sparse germination.  At the Yekaterinino and Ustimovka Stations, drought continued throughout the Summer, resulting in a small seed yield from the majority of the accessions.  Weather conditions at the Kuban Station were equally unfavorable for all crops, including the maize and castor bean fields, where isolated plots of hemp were placed.  The maize failed to develop cobs and thus was harvested early for green fodder, which necessitated premature harvesting of the hemp plants.  The castor bean plants were very low (less than 1 m) and did not conceal the hemp plants, therefore they were stolen.

Table 1.  Results of 1996 VIR/IHA Cannabis Germplasm reproduction at Povlovsk Experimental Station of the VIR

        At the Pavlovsk Station, Spring was dry and the Summer was wet and cold.  The hemp plants were unusually small and low-yielding.   Fifteen accessions of Swedish and Central Russian types were planted in isolated plots (Table 1).  One accession failed to germinate, two were cut prematurely, and 12 were reproduced, including four which yielded less than 100 g of seed.

Table 2.  Results of 1996 VIR/IHA Cannabis Germplasm reproduction at Yekaterinino Experimental Station of the VIR

        On isolated plots of Yekaterinino Station, 20 accessions of Central Russian and southern-ripening types from Germany and the Ukraine were planted (Table 2).  Due to the dry weather, two accessions did not germinate at all, while most of them germinated sparsely.  Therefore, ten accessions produced less than 100 g, while four accessions were stolen and 14 accessions were reproduced successfully.

Table 3.  Results of 1996 VIR/IHA Cannabis Germplasm reproduction at Kuban Experimental Station of the VIR

        At the Kuban Experiment Station, 20 accessions were planted.  They and were mostly southern-ripening types of Chinese origin and also included accessions from Armenia, Italy, Russia, Turkey and the Ukraine (Table 3).  Due to sparse germination and premature harvesting, six accessions produced less than 100 g of seed, one sample failed to germinate, two samples were damaged during chemical weed control for maize, and four accessions were stolen.  Thirteen accessions were reproduced in total.

Table 4.  Results of 1996 VIR/IHA Cannabis Germplasm reproduction at Ustimovka Experimental Station of the VIR

        Thirty-one southern-type accessions were planted at the Ustimovka Experiment Station.  They were mostly of Yugoslavian and Hungarian origin, with additional accessions from Armenia, China, Italy, Turkey, Russia and the Ukraine (Table 4).  Due to dry weather conditions, two accessions failed to germinate and 15 produced sparse plants, which resulted in seed reproductions of less than 100 g.  Twenty-nine accessions were reproduced.

Table 5.  Results of 1996 VIR/IHA Cannabis Germplasm reproduction at the Penza Research Institute of Agriculture

        At the Penza Institute of Agriculture, five hemp accessions belonging to the Central Russian group were planted on isolated plots.   Also included were southern-ripening types from France and Germany (Table 5).   Accession No. 58 from Russia was cut and stolen and only a few plants survived.   Two accessions showed sparse germination and produced seed less than 100 g of seed.   Five accessions were maintained.

Table 6.  Results of 1996 VIR/IHA Cannabis Germplasm reproduction at the Donsky Branch of the All-Russian Institute of Oil-Bearing Crops

        The reproductions on isolated plots at the Donskoy Branch of the Institute of Oil-bearing Crops were not successful.  All five Russian and Ukrainian accessions planted (Table 6) were stolen.  One plant (No. 131 from Ukraine) was by chance preserved, but produced a small number of seeds.  This experience will mean that we must discontinue cooperation with this station.

Table 7.  Results of 1996 VIR/IHA Cannabis Germplasm reproduction at green house conditions Pushkin laboratories of VIR (monoecious accessions)

        In the Pushkin Laboratories glasshouses, 16 accessions of monoecious hemp were maintained in isolated boxes (Table 7).  Regular inspection and elimination of dioecious individuals was performed.  Due to the small number of plants, the seed yield here was insignificant.  However, this experiment shows that using glasshouses for hemp maintenance may be quite helpful.  It is especially convenient for the multiplication of monoecious hemp samples, but late-ripening southern forms can also be reproduced here.

Conclusions
        In 1996, within the framework of the VIR/IHA Cannabis Germplasm Preservation Project, a total of 96 hemp accessions were planted on isolated plots.  Seventy-four accessions yielded fresh reproductions.  In addition, 16 accessions of monoecious hemp were grown in glasshouses and although the yield of these seeds was small, they represent valuable planting materials due to diligent elimination of dioecious plants.  There was an attempt to enlarge the number of accessions maintained by expanding the number of participating stations.  Our experience showed that the Penza Institute deserves inclusion in the list of participants, but further work with Donskoy Branch of the Institute of Oil-bearing Crops seems futile.
        The remaining reproductions should be completed in 1997, but a great deal of work will be required.  More than 120 accessions in the VIR collection must be reproduced because the germplasm dates back to 1989-1991, and many of these old seeds may have lost viability.

References

• Lemeshev, N., L. Rumyantseva and R. C. Clarke  1994.  Maintenance of Cannabis germplasm in the Vavilov Research Institute Gene Bank - 1993.  Journal of the International Hemp Association 1(1): 1, 3-5.
• Lemeshev, N., L. Rumyantseva and R. C. Clarke  1995.  Maintenance of hemp (Cannabis sativa L.) germplasm accessioned in the Vavilov Research Institute Gene Bank - 1994.   Journal of the International Hemp Association 2(1): 10-13.
• Kutuzova, S., L. Rumyantseva and R. C. Clarke  1996.  Maintenance of Cannabis germplasm in the Vavilov Research Institute Gene Bank - 1995.  Journal of the International Hemp Association 3(1): 10-12.